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blunt end - filling, polishing (Aug/01/2007 )

hi

if I cut with HIndIII
I'll generat a fragment like below

A/AGCT T
T TCGA/A

5'---A] AGCTT----3'
3'---TTCGA [A----5'

what is the best way to make it blunt.
filling polishing.

filling will give me that:correct?

5'---AAGCT] [AGCTT----3'
3'---TTCGA] [TCGAA----5'


and polishing will give me this: correct?

5'---A] [T----3'
3'---T] [A----5'


SO if this is correct what enzyme do that

thanks

-ulujm-

I always use Pfu polymerase to blunt sticky ends. In this case, this will fill in and give you the sequence you describe.

-dpo-